Gene silencing with siRNA was performed as described (23 (link), 30 (link), 49 (link)). In brief, to deplete α1B/D-ARs from the cell surface, cells were incubated in Nunc six-well plates (Thermo Fisher Scientific) for 3 d in Accell transfection media (GE Dharmacon) with α1B-AR, α1D-AR, or NT (negative control) siRNA (GE Dharmacon) at a concentration of 1 µM. On day 3, cells were centrifuged at 300 × g for 5 min and resuspended in RPMI 1640 (Sigma) supplemented with 10% HyClone FBS from Cytiva (Marlborough, MA), 100 μg/mL penicillin from Invitrogen (Waltham, MA), and 100 μg/mL streptomycin (Invitrogen). Cells were utilized on day 4 for experimentation.
Nunc six well plates
Manufactured by Thermo Fisher Scientific
Sourced in New Zealand
Nunc six-well plates are a type of cell culture plate designed for in vitro experiments. The plates feature six individual wells, each providing a separate compartment for cell growth and analysis. The plates are made of tissue culture-treated polystyrene, providing a suitable surface for cell attachment and proliferation.
Automatically generated - may contain errors
4 protocols using nunc six well plates
1
Gene Silencing with siRNA Depletion
2
Glucose Deprivation Phospho-Tyrosine Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LN18 parental cells were grown in DMEM without phenol red and supplemented with 5% FBS in a Nunc six‐well plates (Thermo Fisher Scientific) for 2 days. Cells were then treated with chemical inhibitors 1 h before glucose deprivation. Proteins were then harvested 2 h after glucose starvation. Phospho‐tyrosine levels were probed with phospho‐tyrosine (4G10) Mouse mAb (Cat. No. 96215) and quantified using ImageJ.
3
Colony Formation Assay for MCF-7 and AHR Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MCF-7 and AHR100 cells were plated at a density of 2000 cells per well in NUNC six‐well plates (Thermo Fisher Scientific, Rockford, IL) and treated with either DMSO or AF for 24 h, then allowed to grow for 2 weeks before the cells were fixed with 10% formalin and stained with crystal violet solution. Colonies were imaged and counted using the open-source image processing software ImageJ (National Institutes of Health, Bethesda, MD) for colony formation analysis. Use of this software enabled a non-biased assessment of the colony number.
4
Depletion of AVPR1A from Cell Surface
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Gene silencing with siRNA was performed as described previously (14 (link), 15 (link)). In brief, to deplete AVPR1A from the cell surface, cells were incubated in Nunc six-well plates (Thermo Fisher Scientific) for 3 d in Accell transfection media (GE Dharmacon) with AVPR1A or NT (negative control) siRNA (GE Dharmacon) at a concentration of 1 μM. On day 3, cells were centrifuged at 300g for 5 min and resuspended in RPMI 1640 (Sigma-Aldrich) supplemented with 10% HyClone FBS from Cytiva, 100 μg/ml penicillin from Invitrogen (Waltham, MA), and 100 μg/ml streptomycin (Invitrogen). Cells were used on day 4 for experimentation.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!